Ohno Hirohisa, Osada Eriko, Saito Hirohide
Laboratory of Gene Biodynamics, Graduate School of Biostudies, Kyoto University, Kitashirakawa Oiwake-cho, Sakyo-ku, Kyoto, 606-8502, Japan.
Methods Mol Biol. 2015;1297:197-211. doi: 10.1007/978-1-4939-2562-9_14.
The use of RNA-protein interaction motifs (RNP motifs) to design and build nanoscale objects has the potential to expand the field of RNA nanotechnology. In principle, RNP motifs can be integrated easily into RNA nano objects, providing an alternative technique to increase the functional and structural complexities of the RNA. Investigating the design principles of RNP nanostructures will enable the construction of highly sophisticated biomacromolecular complexes such as ribosomes from scratch. As an initial step towards this goal, we designed and constructed triangular-like nanostructures by employing box C/D kink-turn (K-turn)-L7Ae RNP motifs. We showed that the K-turn RNA and the ribosomal protein L7Ae could form a nanostructure shaped like an equilateral triangle that consists of the three proteins attached to the tips of the RNA scaffold. The construction of the complex depends on L7Ae binding to the K-turn motifs in the RNA. The RNP motif allows the RNA to bend by approximately 60° at three positions to form a nanoscale triangle. Functional RNP triangles with desired protein modules at the three tips can be constructed in a modular manner. Here, we describe how to design, construct, and evaluate the RNP nanostructures.
利用RNA-蛋白质相互作用基序（RNP基序）来设计和构建纳米级物体，有可能拓展RNA纳米技术领域。原则上，RNP基序能够轻松整合到RNA纳米物体中，为增加RNA的功能和结构复杂性提供一种替代技术。研究RNP纳米结构的设计原则将使从头构建诸如核糖体等高度复杂的生物大分子复合物成为可能。作为朝着这个目标迈出的第一步，我们通过采用盒C/D扭结转角（K转角）-L7Ae RNP基序设计并构建了三角形样纳米结构。我们表明，K转角RNA和核糖体蛋白L7Ae能够形成一个等边三角形形状的纳米结构，该结构由附着在RNA支架末端的三种蛋白质组成。复合物的构建取决于L7Ae与RNA中的K转角基序结合。RNP基序使RNA在三个位置处弯曲约60°，从而形成一个纳米级三角形。可以以模块化方式构建在三个末端带有所需蛋白质模块的功能性RNP三角形。在此，我们描述如何设计、构建和评估RNP纳米结构。
